Changes in peripheral HCN2 channels during persistent inflammation

ABSTRACT

Nociceptor sensitization following nerve injury or inflammation leads to chronic pain. An increase in the nociceptor hyperpolarization-activated current, I_h, is observed in many models of pathological pain. Pharmacological blockade of I_h prevents the mechanical and thermal hypersensitivity that occurs during pathological pain. Alterations in the Hyperpolarization-activated Cyclic Nucleotide-gated ion channel 2 (HCN2) mediate I_h-dependent thermal and mechanical hyperalgesia. Limited knowledge exists regarding the nature of these changes during chronic inflammatory pain. Modifications in HCN2 expression and post-translational SUMOylation have been observed in the Complete Freund’s Adjuvant (CFA) model of chronic inflammatory pain. Intra-plantar injection of CFA into the rat hindpaw induces unilateral hyperalgesia that is sustained for up to 14 days following injection. The hindpaw is innervated by primary afferents in lumbar DRG, L4-6. Adjustments in HCN2 expression and SUMOylation have been well-documented for L5 DRG during the first 7 days of CFA-induced inflammation. Here, we examine bilateral L4 and L6 DRG at day 1 and day 3 post-CFA. Using L4 and L6 DRG cryosections, HCN2 expression and SUMOylation were measured with immunohistochemistry and proximity ligation assays, respectively. Our findings indicate that intra-plantar injection of CFA elicited a bilateral increase in HCN2 expression in L4 and L6 DRG at day 1, but not day 3, and enhanced HCN2 SUMOylation in ipsilateral L6 DRG at day 1 and day 3. Changes in HCN2 expression and SUMOylation were transient over this time course. Our study suggests that HCN2 is regulated by multiple mechanisms during CFA-induced inflammation.

KEYWORDS:

• HCN2
• CFA
• SUMO
• inflammation
• DRG

Acknowledgments

We would like to thank our labmate Meghyn Welch for her continued support and feedback, and Alex Perez for his expertise in software development and creating a program to aid in data analysis.

Disclosure statement

The authors have no conflicts of interest to declare.

Additional information

Funding

This work was supported by Georgia State University with a Brains and Behavior Seed Grant to [DJB and AZM] and a Brains and Behavior fellowship to [LAF] and by NIH R03NS116327 to [DJB].